title: Hierarchy of lens proteins requiring protection against heat-induced precipitation by the alpha crystallin chaperone.
abstract: Gel filtration of the water-soluble extract from bovine lens yields a group of proteins, emerging between the peaks of beta H and beta L crystallins, which show a considerably greater sensitivity to heat-induced aggregation/precipitation than the far more abundant beta and gamma crystallins. However, the small heat shock protein: alpha crystallin was effective in protecting these trace constituents of the lens from precipitating out of solution at 55 degrees C (measured under the standard conditions in a pH 7.5 buffer containing 50 mM sodium phosphate, 100 mM NaCl, 1 mM EDTA and 0.05% NaN3). Prominent components of the precipitate, formed in the absence of a recombinant alpha B crystallin chaperone could be resolved by one- and two-dimensional electrophoresis. Identification by amino acid sequencing revealed that the heat-sensitive group of lens proteins comprised glyceraldehyde-3-phosphate dehydrogenase (M(r) approximately 39 kDa), enolase (approximately 48 kDa), leucine aminopeptidase (approximately 52 kDa) and aldehyde dehydrogenase (approximately 53 kDa). These findings indicate for the first time that the aggregation of such minor lens constituents could possibly contribute to initiating the process of opacification in the development of cataracts.
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611, USA.
authors.forename: P T
authors.initials: PT
authors.lastname: Velasco
authors.forename: T J
authors.initials: TJ
authors.lastname: Lukas
authors.forename: S N
authors.initials: SN
authors.lastname: Murthy
authors.forename: Y
authors.initials: Y
authors.lastname: Duglas-Tabor
authors.forename: D L
authors.initials: DL
authors.lastname: Garland
authors.forename: L
authors.initials: L
authors.lastname: Lorand
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002386:Cataract / Q000209:etiology / Q000378:metabolism; D002417:Cattle; D002449:Cell Aggregation; D011232:Chemical Precipitation; D003459:Crystallins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D007908:Lens, Crystalline / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism*; D008969:Molecular Sequence Data; D011489:Protein Denaturation
chemical_list: D003459:Crystallins; D018832:Molecular Chaperones